scholarly journals Unexpected gene activation following CRISPR‐Cas9‐mediated genome editing

EMBO Reports ◽  
2021 ◽  
Author(s):  
Anna G Manjón ◽  
Simon Linder ◽  
Hans Teunissen ◽  
Anoek Friskes ◽  
Wilbert Zwart ◽  
...  
Keyword(s):  
Genome Editing ◽  
Gene Activation

A split CRISPR–Cpf1 platform for inducible genome editing and gene activation

2019 ◽  
Vol 15 (9) ◽  
pp. 882-888 ◽  
Author(s):  
Yuta Nihongaki ◽  
Takahiro Otabe ◽  
Yoshibumi Ueda ◽  
Moritoshi Sato
Keyword(s):  
Genome Editing ◽  
Gene Activation

scholarly journals A far-red light–inducible CRISPR-Cas12a platform for remote-controlled genome editing and gene activation

2021 ◽  
Vol 7 (50) ◽  
Author(s):  
Xinyi Wang ◽  
Kaili Dong ◽  
Deqiang Kong ◽  
Yang Zhou ◽  
Jianli Yin ◽  
...  
Keyword(s):  
Genome Editing ◽  
Red Light ◽  
Gene Activation

scholarly journals Biology and Application of Genome Editing

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. SCI-22-SCI-22
Author(s):  
Feng Zhang
Keyword(s):  
Genome Editing ◽  
Dna Sequences ◽  
Large Scale ◽  
Gene Knockout ◽  
Gene Activation ◽  
Base Editing ◽  
Cellular Processes ◽  
Equity Ownership ◽  
Rna Knockdown

Precision genome editing, which can be used to alter specific DNA sequences, is a powerful tool for understanding the molecular circuitry underlying cellular processes. Over the past several years, we and others have harnessed microbial CRISPR-Cas systems for use as platforms for a range of genome manipulations, including single and multiplex gene knockout, gene activation, and large-scale screening applications. Recently, we discovered and characterized several novel CRISPR systems that target RNA, including the CRISPR-Cas13 family. We developed a toolbox for RNA modulation based on Cas13, including methods for highly specific RNA knockdown, transcript imaging, and precision base editing. During our initial characterization of Cas13, we observed that Cas13 also exhibits so-called non-specific "collateral" RNase activity in vitro, which we capitalized on to create SHERLOCK, a highly sensitive and specific CRISPR diagnostic platform. We are continuing to refine and extend CRISPR-based technologies as well as explore microbial diversity to find new enzymes and systems that can be adapted for use as molecular biology tools and novel therapeutics. Disclosures Zhang: Arbor Biotechnologies: Consultancy, Equity Ownership; Sherlock Biosciences: Consultancy, Equity Ownership; Pairwise Plants: Consultancy, Equity Ownership; Beam Therapeutics: Consultancy, Equity Ownership; Editas Medicine: Consultancy, Equity Ownership.

scholarly journals Unexpected gene activation following CRISPR-Cas9-mediated genome editing

2021 ◽  
Author(s):  
Anna G Manjon ◽  
Hans Teunissen ◽  
Elzo de Wit ◽  
Rene H Medema
Keyword(s):  
Dna Damage ◽  
Genome Editing ◽  
Target Location ◽  
Gene Activation ◽  
Regulatory Region ◽  
Lentiviral Vectors ◽  
Strand Breaks ◽  
A Genome ◽  
Damage Field ◽  
Genomic Locations

The discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR) and its development as a genome editing tool has revolutionized the field of molecular biology. In the DNA damage field, CRISPR has brought an alternative to induce endogenous double-strand breaks (DSB) at desired genomic locations and study the DNA damage response and its consequences. Many systems for sgRNA delivery have been reported in order to efficiency generate this DSB, including lentiviral vectors. However, some of the consequences of these systems are yet not well understood. Here we report that lentiviral-based sgRNA vectors can integrate into the endogenous genomic target location, leading to undesired activation of the target gene. By generating a DSB in the regulatory region of the ABCB1 gene using a lentiviral sgRNA vector, we can induce the formation of taxol-resistant colonies. We show that these colonies upregulated ABCB1 via integration of the EEF1A1 and the U6 promoters from the sgRNA vector. We believe that this is an unreported CRISPR/Cas9 artefact that researchers need to be aware of when using lentiviral vectors for genome editing.

Conformational characterization of nucleosome structure by Electron Microscopy

1993 ◽  
Vol 51 ◽  
pp. 194-195
Author(s):  
Gregory J. Czarnota
Keyword(s):  
Electron Microscopy ◽  
Structural Changes ◽  
Physiological State ◽  
Gene Activation ◽  
Three Dimensional ◽  
Macromolecular Complex ◽  
Ultrastructural Studies ◽  
Ionic Environment ◽  
Nucleosome Structure ◽  
Dimensional Reconstruction

Chromatin structure at the fundamental level of the nucleosome is important in vital cellular processes. Recent biochemical and genetic analyses show that nucleosome structure and structural changes are very active participants in gene expression, facilitating or inhibiting transcription and reflecting the physiological state of the cell. Structural states and transitions for this macromolecular complex, composed of DNA wound about a heterotypic octamer of variously modified histone proteins, have been measured by physico-chemical techniques and by enzyme-accessibility and are recognized to occur with various post-translational modifications, gene activation, transformation and with ionic-environment. In spite of studies which indicate various forms of nucleosome structure, all current x-ray and neutron diffraction studies have consistently resulted in only one structure, suggestive of a static conformation. In contrast, two-dimensional electron microscopy studies and three-dimensional reconstruction techniques have yielded different structures. These fundamental differences between EM and other ultrastructural studies have created a long standing quandary, which I have addressed and resolved using spectroscopic electron microscopy and statistical analyses of nucleosome images in a study of nucleosome structure with ionic environment.

Genome editing: Wollen wir das wirklich?

2017 ◽  
Vol 6 (3) ◽  
pp. 162-162
Author(s):  
Liane Kaufmann ◽  
Michael von Aster
Keyword(s):  
Genome Editing

Genome editing (CRISPR-Cas9) to identify and characterise functional variants determining metformin response

2018 ◽  
Author(s):  
M Keller ◽  
J Dalla-Riva ◽  
A Kurbasic ◽  
M Al-Majdoub ◽  
P Spegel ◽  
...  
Keyword(s):  
Genome Editing ◽  
Functional Variants

GENE ACTIVATION IN MAMMARY CELLS

1972 ◽  
Vol 71 (2_Suppla) ◽  
pp. S346-S368 ◽  
Author(s):  
Roger W. Turkington ◽  
Nobuyuki Kadohama
Keyword(s):  
Cell Differentiation ◽  
Gene Transcription ◽  
Hormonal Regulation ◽  
Gene Activation ◽  
Milk Proteins ◽  
Mouse Mammary Gland ◽  
Nuclear Proteins ◽  
Mammary Cells ◽  
Alveolar Cells ◽  
Mammary Cell

ABSTRACT Hormonal activation of gene transcription has been studied in a model system, the mouse mammary gland in organ culture. Transcriptive activity is stimulated in mammary stem cells by insulin, and in mammary alveolar cells by prolactin and insulin. Studies on the template requirement for expression of the genes for milk proteins demonstrate that DNA methylation has an obligatory dependence upon DNA synthesis, but is otherwise independent from hormonal regulation of mammary cell differentiation. Incorporation of 5-bromo-2′deoxyuridine into DNA selectively inhibits expression of the genes for specific milk proteins. Undifferentiated mammary cells activate the synthesis of specific acidic nuclear proteins when stimulated by insulin. Several of these induced acidic nuclear proteins are undetectable in unstimulated undifferentiated cells, but appear to be characteristic components of the nuclei of differentiated cells. These results indicate that mammary cell differentiation is associated with a change in acidic nuclear proteins, and they provide evidence to support the concept that acidic nuclear proteins may be involved in the regulation of gene transcription and of mammary cell differentiation.

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